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Methods for rapid screening and isolation of bacteria producing acidic lipase: feasibility studies and novel activity assay protocols

机译:快速筛选和分离产生酸性脂肪酶细菌的方法:可行性研究和新的活性测定方案

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摘要

Acidic lipase finds its commercial values in medical applications and bioremediation of food wastes. In this work, approaches for rapid screening of lipase-producing bacteria were developed and the feasibility assessment of the screening methods was performed. From food waste samples, the proposed screening procedures allowed isolation of sixteen pure bacterial strains expressing higher lipase activity at acidic pH (pH 6.0) than at alkaline pH (pH 9.0). To enhance the accuracy of lipase activity determination under acidic conditions, a novel assay procedure was also developed by deactivating lipase activity by microwave treatment prior to back titration. This additional step could minimize interferences arising from residual lipase activity during conventional direct back-titration methods in measuring lipase activity at acidic pH. Using the four strategies proposed in this work, the best acidic-lipase-producing isolate was obtained by strategy C (SSC) and was identified as iAeromonas sp. C14, displaying an optimal lipase activity of 0.7 U/ml at an acidic pH of 6.0.
机译:酸性脂肪酶在医疗应用和食物垃圾的生物修复中具有商业价值。在这项工作中,开发了快速筛选产脂肪酶细菌的方法,并对筛选方法进行了可行性评估。从食物垃圾样品中,建议的筛选程序可以分离出16种在酸性pH(pH 6.0)下比在碱性pH(pH 9.0)下表达更高脂肪酶活性的纯细菌菌株。为了提高在酸性条件下脂肪酶活性测定的准确性,还开发了一种新的测定方法,该方法通过在反滴定之前通过微波处理使脂肪酶活性失活。在常规的直接反滴定方法中,在酸性pH值下测量脂肪酶活性时,该附加步骤可以最大程度地减少由残留脂肪酶活性引起的干扰。使用这项工作中提出的四种策略,通过策略C(SSC)获得了最佳的酸性脂肪酶生产菌,并被鉴定为iAeromonas sp。 C14,在6.0的酸性pH下显示0.7 U / ml的最佳脂肪酶活性。

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